Comparative effect of Ovysynch Protocol and dietary Trigonella foenum graceum seed supplementation on hormonal, biochemical and antioxidant profiles in postpartum anestrum buffaloes

Introduction

Postpartum anestrum remains one of the major reproductive constraints limiting productivity in dairy buffaloes. In India and other sub-tropical areas, postpartum anestrum in buffaloes may be due to several factors like negative energy balance, lack of antioxidants, hormonal imbalance, parity, anaemia etc. (Misra et al., 2023) [11] . The prolonged anestrus period directly reduces calving rate, increases calving interval and contributes to economic losses in milk production and efficient reproduction. To combat postpartum anestrum in buffaloes, several hormonal therapies were applied that can restore ovarian cyclicity like GnRH, progesterone, insulin, PMSG (CIRD+PRID). The most effective and conventional applied hormonal protocol used in cattle and buffaloes is Ovysynch (Brahmanand et al., 2023) [4] . The protocol uses sequential administration of GnRH and PGF 2α to synchronize follicular waves and induce ovulation, thereby shortening the anestrum period and enabling timed artificial insemination. Though effective, repeated hormonal use raises concerns about cost, residues, consumer perception and long term effects on reproductive physiology. Even prolonged or modified Ovsynch protocols can lead to reduced fertility, lower conception rates, and premature estrus, particularly if implemented without proper veterinary screening. (Sun S et al., 2024) [19] . As an alternate to hormonal therapy, there are several non-hormonal herbal plants like Murraya koenigii (curry takes off), Nigella sativa (kalonji), Abroma augusta (Ulatkambal), Saraca asoca (Ashoka), Trigonella foenum-graecum (Methi), Asparagus recemosus (Shatavari), Bambusa aruninacea, Leptadenia reticulate, Pergulacia daemia, Semecarpus anacardium (Chandra Shekhar Sarswat 2020) [5] . Due to antioxidant and anti-inflammatory compounds present in herbal plants, dietary intake of them will indirectly or directly ameliorate anestrum in postpartum buffaloes and also show other benefits like biochemical, antioxidant and anti-inflammatory effect. Further there are non-toxic. Triogonella foenum graecum seeds contain diosgenin, alkaloids, flavonoids, polyphenols that contribute to antioxidant, antiinflammatory, antidiabetic, hypocholesterolemic activities. (Akhtar et al., 2025) [2] . So, the present study is to compare effect of Ovysynch Protocol and dietary Trigonella foenum graceum seed supplementation on hormonal, biochemical and antioxidant profiles in postpartum anestrum buffaloes.

Materials and methods

A total of sixteen high yielding buffaloes after postpartum anestrum (>90 days) were selected after examination of follicular wave dynamics by routine trans -rectal ultrasonography and were divided into two groups; Group I was treated with Ovysynch protocol (GnRH -PGF 2α -GnRH on 0-7-9 th day respectively) while group II were fed orally with Trigonella foenum graceum seeds powder @ 200gm daily once for 24 days. Blood samples were collected in specified vacutainers at an interval of 0, 8, 16 and 24 th days of feeding. Serum was isolated and analysed for hormonal profile viz Progesterone and 17β-estradiol, biochemical parameters like glucose, protein, albumin, globulin, A/G ratio, total cholesterol, and triglycerides; antioxidant parameters viz Lipid peroxidase, SOD and catalase.

Results and Discussion

The values of serum progesterone (ng/ml) and serum 17βestradiol (pg/ml) were depicted in Table 1. Serum progesterone in Group I revealed significantly (p<0.01) higher than group II on 16 th , 20 th day and Group II showed increased levels on 20 and 24 th day indicating that some animals had showed estrous on these days. This is similar to the findings of Rajkumar et al., 2008 14 . The group II showed estrous in some animals might be due to diosgenin in Trigonella foenum graceum seeds which will promote ovarian folliculogenesis, hormone release, oocyte quality and prevent reproductive aging (Sirotkin et al., 2019) [18] . The mean values of serum 17β-estradiol in Group I were significantly increased on 12 th day whereas in group II the mean values were increased on 16 th day indicating that the animals showed estrous in these days. Hind et al., (2019) [7] observed that on feeding of fenugreek aqueous extract orally, the estrogenic activity was shown in vagina through an increase in epidermal cell number, tendency to keratinization, overexpression of progesterone receptors in the uterine tissue. [10] .

Decreased blood glucose levels may be due to insulin release that effects the peripheral tissue by promoting glucose uptake and metabolism, by attenuating the key enzymes of carbohydrate metabolism or by inhibiting hepatic gluconeogenesis (Saravanan et.al., 2014) [15] . Hepatic enzymes like alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, lactate dehydrogenase and alkaline phosphatase lower the glycogen level thus reduce serum glucose (Kaviarasam et al., 2007) [8] . Increase in serum protein, albumin and globulin is observed in Group II compared to Group I. Similar findings were observed by feeding 200gm of fenugreek seeds in buffaloes by Abo El-Nor et.al., (2007) [1] .; Kriar et.al., 2018 [9] . A/G ratio did not show any significant difference. Serum total cholesterol, triglycerides were reduced in group II compared to group I which might be due to the effect of disogenin in Trigonella foenum graecum seeds. Diosgenin will increase ABCG5/G8 complex expression in liver and intestine which inhibit absorption of intestinal cholesterol by antagonizing Niemann-Pick C1-Like 1 (NPC1L1) function. (DikkersA et al., 2010) [6] . Further it increases cholesterol 7α-hydroxylase or cytochrome P450 7A1 (CYP7A1), carboxylesterase-1 (CES-1), and scavenger receptor class B type I (SRB1) levels in liver resulting in promoting reverse cholesterol transport, inhibit cholesterol absorption and eliminate cholesterol (Shi et al., 2021) [16] . Diosgenin will upregulate sirtuin 6 expression and fatty acid transporters inhibiting liver from fatty acid absorption (Nie K et al., 2023) [13] . Diosgenin enhancing biliary secretion and modify bile acid synthesis by regulating gut microbiome. It can decrease serum triglycerides and phospholipids and increased serum HDL concentration (Naskar and Mishra, 2017) [13] . Serum lipid peroxidase was reduced and superoxide dismutase activity, catalase activity increased in group II compared to group I. This is similar to the findings of Singaravelu et al., (2018) [17] .

where the fenugreek treated animals showed increase in catalase, SOD and GSH compared to control positive group and significant decrease in TBARS value in treated group. The flavonoids and polyphenols in Trigonella foenum graecum seeds will accelerate antioxidant enzymes like catalase, SOD, glutathione peroxidase, glutathione reductase in the liver, skeletal muscle, cardiac muscle and brain (Baquer et al., 2011) [3] .

Conclusion

From the above study, postpartum anestrum in buffaloes on treatment with both conventional hormonal protocol and feeding with Trigonella foenum greacum seed powder showed estrous symptoms. Though Group I showed estrous earlier compared to group II, but in case of the biochemical profile and antioxidant profile, group II (FS treatment) has more beneficial effect compared to conventional hormonal protocol along with exhibiting of estrous cycle.

References

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